title: Effect of cell culture medium components on color of formulated monoclonal antibody drug substance.
abstract: As the industry moves toward subcutaneous delivery as a preferred route of drug administration, high drug substance concentrations are becoming the norm for monoclonal antibodies. At such high concentrations, the drug substance may display a more intense color than at the historically lower concentrations. The effect of process conditions and/or changes on color is more readily observed in the higher color, high concentration formulations. Since color is a product quality attribute that needs to be controlled, it is useful to study the impact of process conditions and/or modifications on color. This manuscript summarizes cell culture experiments and reports on findings regarding the effect of various media components that contribute to drug substance color for a specific monoclonal antibody. In this work, lower drug substance color was achieved via optimization of the cell culture medium. Specifically, lowering the concentrations of B-vitamins in the cell culture medium has the effect of reducing color intensity by as much as 25%. In addition, decreasing concentration of iron was also directly correlated color intensity decrease of as much as 37%. It was also shown that the color of the drug substance directly correlates with increased acidic variants, especially when increased iron levels cause increased color. Potential mechanisms that could lead to antibody coloration are briefly discussed.
authors.affiliation: Late Stage Cell Culture, Genentech Inc., South San Francisco, CA, 94080.
authors.forename: Natarajan
authors.initials: N
authors.lastname: Vijayasankaran
authors.forename: Sharat
authors.initials: S
authors.lastname: Varma
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Melissa
authors.initials: M
authors.lastname: Mun
authors.forename: Silvana
authors.initials: S
authors.lastname: Arevalo
authors.forename: Martin
authors.initials: M
authors.lastname: Gawlitzek
authors.forename: Trevor
authors.initials: T
authors.lastname: Swartz
authors.forename: Amy
authors.initials: A
authors.lastname: Lim
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.forename: Boyan
authors.initials: B
authors.lastname: Zhang
authors.forename: Steve
authors.initials: S
authors.lastname: Meier
authors.forename: Robert
authors.initials: R
authors.lastname: Kiss
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D019149:Bioreactors; D016466:CHO Cells; D018929:Cell Culture Techniques; D002852:Chromatography, Ion Exchange; D003116:Color; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D005492:Folic Acid / Q000494:pharmacology; D007501:Iron / Q000494:pharmacology; D011730:Pyridoxal / Q000494:pharmacology; D011736:Pyridoxine / Q000494:pharmacology; D012256:Riboflavin / Q000494:pharmacology; D014805:Vitamin B 12 / Q000494:pharmacology; D014803:Vitamin B Complex / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D003470:Culture Media; D014803:Vitamin B Complex; D011730:Pyridoxal; D005492:Folic Acid; D007501:Iron; D011736:Pyridoxine; D014805:Vitamin B 12; D012256:Riboflavin
keywords: B12; B2; B6; B9; CHO; antibody; cell culture; fed-batch; ferrous sulfate; iron; medium optimization; protein color